大黄鱼（<em>Larimichthys crocea</em>）IL-10基因的克隆与表达分析

郑维兵; 慕鹏飞; 丁连帅; 杨思司; 敖敬群; 艾春香; 陈新华

doi:10.3969/j.issn.0253-4193.2017.04.005

大黄鱼（Larimichthys crocea）IL-10基因的克隆与表达分析

doi: 10.3969/j.issn.0253-4193.2017.04.005

1.
国家海洋局第三海洋研究所海洋生物遗传与资源重点实验室, 福建厦门 361005
2.
厦门大学海洋与地球学院, 福建厦门 361005

基金项目: 国家自然科学基金资助项目（31001131 ，31372556）；厦门南方海洋研究中心资助项目（13GZP002NF08，14CZP049SF02）。

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出版历程
- 收稿日期: 2016-07-01
- 修回日期: 2016-07-31

Cloning and expression analysis of large yellow croaker (Larimichthys rocea) IL-10 gene

1.
Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China
2.
College of Ocean and Earth Sciences, Xiamen University, Xiamen 361005, China

摘要

摘要:
白细胞介素10（Interleukin 10，IL-10）是一种抗炎细胞因子，可以抑制机体免疫反应。本研究经过分析大黄鱼基因组数据库发现了IL-10同源基因，并对其cDNA编码区序列和基因组DNA序列进行了克隆分析。大黄鱼IL-10（LycIL-10）基因由5个外显子和4个内含子构成，其序列全长1 869 bp，其开放阅读框（ORF）长555 bp，编码184个氨基酸，其N端的22个氨基酸残基为预测的信号肽，成熟肽由162个氨基酸残基组成，包含了脊椎动物IL-10标志性保守序列。LycIL-10的氨基酸序列同其他已知物种的IL-10氨基酸序列的一致性为26.49%~77.01%。Real-time PCR分析发现LycIL-10在检测的组织中为组成型表达，在脾脏和肌肉中转录水平相对较高。三联灭活细菌疫苗和聚肌苷酸胞苷酸（poly（I：C））刺激后，大黄鱼头肾和脾脏中LycIL-10 mRNA的转录水平会显著升高，表明LycIL-10可能参与抑制大黄鱼由细菌和病毒引起的炎症反应。
- 大黄鱼 /
- 白细胞介素10 /
- 分子克隆 /
- 实时定量PCR /
- 表达分析
Abstract:
Interleukin-10 (IL-10) is a central anti-inflammatory cytokine that demonstrates immunosuppressive function. In this research, an interleukin-10 (IL-10) homologue has been identified by analyzed the genomic data of large yellow croaker (Larimichthys crocea). Subsequently, the open reading frame (ORF) of cDNA and the genomic DNA sequences have been cloned and analyzed. The genomic DNA of large yellow croaker IL-10(LycIL-10) consisted of 1 869 bp that contains five exons and four introns, sharing the same organization with mammalian IL-10 genes. The open reading frame of LycIL-10 consisted 555 bp that give a predicted 184 amino acid IL-10 molecule. It contains a predicted signal peptide of 22 amino acids in the N-terminal and a mature peptide of 162 amino acids, which contained the vertebrate IL-10 family signature. The LycIL-10 exhibits a conserved IL-10 motif signature and shares 26.49%-77.01% amino acid sequence identity with other known IL-10. Real-time PCR analysis showed that LycIL-10 was constitutively expressed in all tissues tested, especially in spleen and muscle. The LycIL-10 mRNA transcription level could significantly increase in the spleen and head kidney of large yellow croaker after trivalent inactivated vaccines or poly(I:C) stimulated, indicating that LycIL-10 may be involved in suppressing the inflammatory induced by bacterium or virus in the large yellow croaker.
- large yellow croaker /
- interleukin-10 /
- molecule cloning /
- real-time PCR /
- expression analysis

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参考文献(51)

Moore K W, de Waal Malefyt R, Coffman R L, et al. Interleukin-10 and the interleukin-10 receptor[J]. Annual Review of Immunology, 2001, 19(1): 683-765.

Fiorentino D F, Bond M W, Mosmann T R. Two types of mouse T helper cell. IV. Th2 clones secrete a factor that inhibits cytokine production by Th1 clones[J]. The Journal of Experimental Medicine, 1989, 170(6): 2081-2095.

Thomson A W, Lotze M T. The Cytokine Handbook, Two-volume Set[M]. 4th ed. London: Gulf Professional Publishing, 2003.

Bogdan C, Vodovotz Y, Nathan C. Macrophage deactivation by interleukin 10[J]. The Journal of Experimental Medicine, 1991, 174(6): 1549-1555.

Ding L, Shevach E M. IL-10 inhibits mitogen-induced T cell proliferation by selectively inhibiting macrophage costimulatory function[J]. The Journal of Immunology, 1992, 148(10): 3133-3139.

Fiorentino D F, Zlotnik A, Mosmann T R, et al. IL-10 inhibits cytokine production by activated macrophages[J]. The Journal of Immunology, 1991, 147(11): 3815-3822.

Fiorentino D F, Zlotnik A, Vieira P, et al. IL-10 acts on the antigen-presenting cell to inhibit cytokine production by Th1 cells[J]. The Journal of Immunology, 1991, 146(10): 3444-3451.

Aste-Amezaga M, Ma Xiaojing, Sartori A, et al. Molecular mechanisms of the induction of IL-12 and its inhibition by IL-10[J]. The Journal of Immunology, 1998, 160(12): 5936-5944.

Gazzinelli R T, Oswald I P, James S L, et al. IL-10 inhibits parasite killing and nitrogen oxide production by IFN-gamma-activated macrophages[J]. The Journal of Immunology, 1992, 148(6): 1792-1796.

Mertz P M, DeWitt D L, Stetler-Stevenson W G, et al. Interleukin 10 suppression of monocyte prostaglandin H synthase-2. Mechanism of inhibition of prostaglandin-dependent matrix metalloproteinase production[J]. The Journal of Biological Chemistry, 1994, 269(33): 21322-21329.

Niiro H, Otsuka T, Kuga S, et al. IL-10 inhibits prostaglandin E₂ production by lipopolysaccharide-stimulated monocytes[J]. International Immunology, 1994, 6(4): 661-664.

de Waal Malefyt R, Yssel H, de Vries J E. Direct effects of IL-10 on subsets of human CD4⁺ T cell clones and resting T cells. Specific inhibition of IL-2 production and proliferation[J]. The Journal of Immunology, 1993, 150(11): 4754-4765.

Groux H, Bigler M, de Vries J E, et al. Inhibitory and stimulatory effects of IL-10 on human CD8⁺ T cells[J]. The Journal of Immunology, 1998, 160(7): 3188-3193.

Vieira P, de Waal-Malefyt R, Dang M N, et al. Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: homology to Epstein-Barr virus open reading frame BCRFI[J]. Proceedings of the National Academy of Sciences of the United States of America, 1991, 88(4): 1172-1176.

Villinger F, Brar S S, Mayne A, et al. Comparative sequence analysis of cytokine genes from human and nonhuman primates[J]. The Journal of Immunology, 1995, 155(8): 3946-3954.

Hash S M, Brown W C, Rice-Ficht A C. Characterization of a cDNA encoding bovine interleukin 10: kinetics of expression in bovine lymphocytes[J]. Gene, 1994, 139(2): 257-261.

Syto R, Murgolo N J, Braswell E H, et al. Structural and biological stability of the human interleukin 10 homodimer[J]. Biochemistry, 1998, 37(48): 16943-16951.

Walter M R, Nagabhushan T L. Crystal structure of interleukin 10 reveals an interferon gamma-like fold[J]. Biochemistry, 1995, 34(38): 12118-12125.

Yoon S I, Logsdon N J, Sheikh F, et al. Conformational changes mediate interleukin-10 receptor 2 (IL-10R2) binding to IL-10 and assembly of the signaling complex[J]. The Journal of Biological Chemistry, 2006, 281(46): 35088-35096.

Liu Y, Wei S H, Ho A S, et al. Expression cloning and characterization of a human IL-10 receptor[J]. The Journal of Immunology, 1994, 152(4): 1821-1829.

Tan J C, Braun S, Rong Hong, et al. Characterization of recombinant extracellular domain of human interleukin-10 receptor[J]. The Journal of Biological Chemistry, 1995, 270(21): 12906-12911.

Tan J C, Indelicato S R, Narula S K, et al. Characterization of interleukin-10 receptors on human and mouse cells[J]. The Journal of Biological Chemistry, 1993, 268(28): 21053-21059.

Wehinger J, Gouilleux F, Groner B, et al. IL-10 induces DNA binding activity of three STAT proteins (Stat1, Stat3, and Stat5) and their distinct combinatorial assembly in the promoters of selected genes[J]. FEBS Letters, 1996, 394(3): 365-370.

Auernhammer C J, Bousquet C, Melmed S. Autoregulation of pituitary corticotroph SOCS-3 expression: characterization of the murine SOCS-3 promoter[J]. Proceedings of the National Academy of Sciences of the United States of America, 1999, 96(12): 6964-6969.

Berlato C, Cassatella M A, Kinjyo I, et al. Involvement of suppressor of cytokine signaling-3 as a mediator of the inhibitory effects of IL-10 on lipopolysaccharide-induced macrophage activation[J]. The Journal of Immunology, 2002, 168(12): 6404-6411.

Qin Hongwei, Wilson C A, Roberts K L, et al. IL-10 inhibits lipopolysaccharide-induced CD40 gene expression through induction of suppressor of cytokine signaling-3[J]. The Journal of Immunology, 2006, 177(11): 7761-7771.

Hsu D H, de Waal Malefyt R, Fiorentino D F, et al. Expression of interleukin-10 activity by Epstein-Barr virus protein BCRF1[J]. Science, 1990, 250(4982): 830-832.

Kotenko S V, Saccani S, Izotova L S, et al. Human cytomegalovirus harbors its own unique IL-10 homolog (cmvIL-10)[J]. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(4): 1695-1700.

Zou J, Clark M S, Secombes C J. Characterisation, expression and promoter analysis of an interleukin 10 homologue in the puffer fish, Fugu rubripes[J]. Immunogenetics, 2003, 55(5): 325-335.

Savan R, Igawa D, Sakai M. Cloning, characterization and expression analysis of interleukin-10 from the common carp, Cyprinus carpio L.[J]. European Journal of Biochemistry, 2003, 270(23): 4647-4654.

Inoue Y, Kamota S, Ito K, et al. Molecular cloning and expression analysis of rainbow trout (Oncorhynchus mykiss) interleukin-10 cDNAs[J]. Fish & Shellfish Immunology, 2005, 18(4): 335-344.

肖凡书, 昌鸣先, 孙军, 等. 鲢(Hypophthalmichthys molitrix)IL-10基因的克隆及表达分析[J]. 自然科学进展, 2006, 16(2): 183-189. Xiao Fanshu, Chang Mingxian, Sun Jun, et al. Cloning and expression analysis of interleukin-10 from the silver carp, Hypophthalmichthys molitrix[J]. Progress in Natural Science, 2006, 16(2): 183-189.

Zhang Dianchang, Shao Yanqing, Huang Yanqin, et al. Cloning, characterization and expression analysis of interleukin-10 from the zebrafish (Danio rerion)[J]. BMB Reports, 2005, 38(5): 571-576.

Pinto R D, Nascimento D S, Reis M I R, et al. Molecular characterization, 3D modelling and expression analysis of sea bass (Dicentrarchus labrax L.) interleukin-10[J]. Molecular Immunology, 2007, 44(8): 2056-2065.

Seppola M, Larsen A N, Steiro K, et al. Characterisation and expression analysis of the interleukin genes, IL-1β, IL-8 and IL-10, in Atlantic cod (Gadus morhua L.)[J]. Molecular Immunology, 2008, 45(4): 887-897.

Grayfer L, Hodgkinson J W, Hitchen S J, et al. Characterization and functional analysis of goldfish (Carassius auratus L.) interleukin-10[J]. Molecular Immunology, 2011, 48(4): 563-571.

Harun N O, Costa M M, Secombes C J, et al. Sequencing of a second interleukin-10 gene in rainbow trout Oncorhynchus mykiss and comparative investigation of the expression and modulation of the paralogues in vitro and in vivo[J]. Fish & Shellfish Immunology, 2011, 31(1): 107-117.

Piazzon M C, Savelkoul H F J, Pietretti D, et al. Carp IL10 has anti-inflammatory activities on phagocytes, promotes proliferation of memory T cells, and regulates B cell differentiation and antibody secretion[J]. The Journal of Immunology, 2015, 194(1): 187-199.

van Beurden S J, Forlenza M, Westphal A H, et al. The alloherpesviral counterparts of interleukin 10 in European eel and common carp[J]. Fish & Shellfish Immunology, 2011, 31(6): 1211-1217.

Sunarto A, Liongue C, McColl K A, et al. Koi herpesvirus encodes and expresses a functional interleukin-10[J]. Journal of Virology, 2012, 86(21): 11512-11520.

Piazzon M C, Wentzel A S, Tijhaar E J, et al. Cyprinid herpesvirus 3 Il10 inhibits inflammatory activities of carp macrophages and promotes proliferation of IgM⁺ B cells and memory T cells in a manner similar to carp Il10[J]. The Journal of Immunology, 2015, 195(8): 3694-3704.

Chen Ruanni, Su Yongquan, Wang Jun, et al. Molecular characterization and expression analysis of interferon-gamma in the large yellow croaker Larimichthys crocea[J]. Fish & Shellfish Immunology, 2015, 46(2): 596-602.

Wu Jun, Shi Yuhong, Zhang Xueheng, et al. Molecular characterization of an IL-1β gene from the large yellow croaker (Larimichthys crocea) and its effect on fish defense against Vibrio alginolyticus infection[J]. Zoological Research, 2015, 36(3): 133-141.

Mu Yinnan, Wang Kunru, Ao Jingqun, et al. Molecular characterization and biological effects of a CXCL8 homologue in large yellow croaker (Larimichthys crocea)[J]. Fish & Shellfish Immunology, 2015, 44(2): 462-470.

Ding Yang, Ao Jingqun, Ai Chunxiang, et al. Molecular and functional identification of three interleukin-17A/F (IL-17A/F) homologues in large yellow croaker (Larimichthys crocea)[J]. Developmental & Comparative Immunology, 2016, 55: 221-232.

Zhu Qian, Li Chan, Yu Zhenxing, et al. Molecular and immune response characterizations of IL-6 in large yellow croaker (Larimichthys crocea)[J]. Fish & Shellfish Immunology, 2016, 50: 263-273.

Ao Jingqun, Mu Yinnan, Xiang Lixin, et al. Genome sequencing of the perciform fish Larimichthys crocea provides insights into molecular and genetic mechanisms of stress adaptation[J]. PLoS Genetics, 2015, 11(4): e1005118.

Zheng Wenbiao, Tian Chen, Chen Xinhua. Molecular characterization of goose-type lysozyme homologue of large yellow croaker and its involvement in immune response induced by trivalent bacterial vaccine as an acute-phase protein[J]. Immunology Letters, 2007, 113(2): 107-116.

Salinas I, Zhang Yong'an, Sunyer J O. Mucosal immunoglobulins and B cells of teleost fish[J]. Developmental & Comparative Immunology, 2011, 35(12): 1346-1365.

Ouyang Wenjun, Rutz S, Crellin N K, et al. Regulation and functions of the IL-10 family of cytokines in inflammation and disease[J]. Annual Review of Immunology, 2011, 29(1): 71-109.

Mu Yinnan, Wan Xiang, Lin Kebing, et al. Liver proteomic analysis of the large yellow croaker (Pseudosciaena crocea) following polyriboinosinic: polyribocytidylic acid induction[J]. Fish Physiology and Biochemistry, 2013, 39(5): 1267-1276.

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